Spectrin-anchored phosphodiesterase 4D4 restricts cAMP from disrupting microtubules and inducing endothelial cell gap formation

Creighton, Judy; Zhu, Bing; Alexeyev, Mikhail; Stevens, Troy

doi:10.1242/jcs.011692

Cited by 51 publications

(60 citation statements)

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“…The influence of tau in axonal development of hippocampal cells (Dawson et al, 2001) might contribute to the proper functioning of specific connexins during electrotonic communication in hippocampal interneurons. Although a role for tau has been demonstrated in the formation of interendothelial cell gap-junctions (Creighton et al, 2008), the link between tau and neuronal gap-junction channels remains to be elucidated.…”

Section: Discussionmentioning

confidence: 99%

Role of tau protein on neocortical and hippocampal oscillatory patterns

et al. 2011

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ABSTRACT:Tau is a neuronal microtubule-associated protein implicated in microtubules stabilization, axonal establishment and elongation during neuronal morphogenesis. Because of its elevated expression in neocortical regions and hippocampus, tau might play a role in sculpting collective neural responses underlying slow and fast brain oscillations and/or long-range synchronization patterns between hippocampus and neocortex. To test this hypothesis, local field potentials were recorded in tau-deficient (tau 2/2 ) and wild-type mice from different neocortical regions and from the hippocampus during spontaneous motor exploratory behavior. We found that tau 2/2 mice showed hippocampal theta slowing and reduced levels of gamma long-range synchronization involving the frontal cortex. We hypothesize that the lack of normal phosphorylated tau during early stages of development might influence the maturation of parvalbumin interneurons affecting the spatiotemporal structure of long-range gamma synchronization. Also, the proper functioning of gap-junction channels might be compromised by the absence of tau in hippocampal networks. Altogether, these results provide novel insights into the functional role of tau protein in the formation of collective neural responses and emergence of neocortical-hippocampal interactions in the mammalian brain. V V C 2010 Wiley-Liss, Inc.

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Section: Discussionmentioning

confidence: 99%

Role of tau protein on neocortical and hippocampal oscillatory patterns

et al. 2011

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“…Rat PMVECs were isolated and cultured in DMEM supplemented with 10% FBS and penicillin/streptomycin using previously described methods (13,34). Cells infected with retrovirus to deliver genes of interest, i.e., PDE4D41-166-green fluorescent protein (GFP), human tau (hTau)-Ser214, hTau-S214A alanine mutation, and vector [GFP or hemagglutinin (HA) tag] controls, were maintained in culture under the same conditions.…”

Section: Methodsmentioning

confidence: 99%

“…Cell extractions in lysis buffer were incubated with antibodies, including selective antibodies for polymerized (SMI62) and depolymerized (SMI61) microtubules (Covance), and EZview Red Protein A Affinity Gel beads (Sigma) at 4°C for 12 h, as previously described (13). The gels were washed with the same incubation buffer and used for Western blot assays.…”

Section: Methodsmentioning

confidence: 99%

“…This protective action of PKA likely reflects compartmentation of the cAMP signal (12,35,37), as near membrane cAMP signals strengthen the cortical actin rim and reduce endothelial cell permeability. In pulmonary microvascular endothelial cells (PMVECs), phosphodiesterase 4D4 (PDE4D4) limits cAMP access to nonneuronal tau (13). Inhibiting PDE4D4 enables cAMP to access the cytosol, where it promotes tau-Ser214 phosphorylation.…”

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Protein kinase A phosphorylation of tau-serine 214 reorganizes microtubules and disrupts the endothelial cell barrier

Zhu

Zhang

Creighton

et al. 2010

American Journal of Physiology-Lung Cellular and Molecular Phys

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compartmentalized to near membrane domains in endothelium, where it strengthens endothelial cell barrier function. Phosphodiesterase 4D4 (PDE4D4) interacts with the spectrin membrane skeleton and prevents cAMP from accessing microtubules. Expression of a dominant-negative PDE4D4 peptide enables cAMP to access microtubules, where it results in phosphorylation of the nonneuronal microtubule-associated protein tau at serine 214. Presently, we sought to determine whether PKA is responsible for tau-Ser214 phosphorylation and furthermore whether PKA phosphorylation of tau-Ser214 is sufficient to reorganize microtubules and induce endothelial cell gaps. In cells expressing the dominant-negative PDE4D4 peptide, forskolin activated transmembrane adenylyl cyclases, increased cAMP, and induced tau-Ser214 phosphorylation that was accompanied by microtubule reorganization. PKA catalytic and regulatory I subunits, but not the regulatory II subunit, coassociated with reorganized microtubules. To determine the functional consequence of tau-Ser214 phosphorylation, wild-type human tau40 and tau40 engineered to possess an alanine point mutation (S214A) were stably expressed in endothelium. In cells expressing the dominant-negative PDE4D4 peptide and tau-S214A, PKA-dependent phosphorylation of both the endogenous and heterologously expressed tau were abolished. Expression of tau-S214A prevented forskolin from depolymerizing microtubules, inducing intercellular gaps, and increasing macromolecular permeability. These findings therefore identify nonneuronal tau as a critical cAMP-responsive microtubule-associated protein that controls microtubule architecture and endothelial cell barrier function. adenylyl cyclase; cytoskeleton; phosphodiesterase ENDOTHELIAL CELLS FORM A RESTRICTIVE barrier that limits fluid, solute, and macromolecule access to the interstitium. The microtubule network plays a key role in establishing endothelial cell shape and hence barrier integrity (17, 41). Whereas stabilizing microtubules enhances endothelial barrier function, disrupting microtubules induces interendothelial cell gap formation that increases permeability (33,47).Tau and other microtubule-associated proteins bind to and stabilize microtubules (3,45). Although tau is a relatively small protein, it shares a conserved 18-amino acid microtubule-binding repeat in common with larger microtubule-associated proteins (3, 44). Tau isoforms vary in their number of repeat domains (20), with the longest tau40 isoform having 4 microtubule-binding repeats at residues 256 -273, 287-304, 318 -335, and 350 -367.Tau40 possesses multiple consensus phosphorylation sites (3, 25); 20% of the tau40 amino acid sequence comprises serine, threonine, and tyrosine residues that can potentially be phosphorylated (25). At least two types of kinase phosphorylation sites exist in tau proteins: proline-direct kinase sites, which possess a threonine-proline (TP)-serine-proline (SP) sequence, and nonproline-directed sites. Most of the TP-SP sites are located in either the NH 2 -or C...

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“…Spectrins have been further related to cellular morphologic changes by their connection to phosphodiesterase and cAM P. [3][4] It is believed that spectrins are also involved in signal transduction, thereby regulating normal cellular function, cellular proliferation, and cell cycles. [5][6][7][8] Importantly, spectrin loss has been shown to impair cell adhesion and cause cell spreading.…”

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Loss of αII and βIII Spectrin Isoforms in Body Fluids is Associated with Metastasis

Wang

Khader

Mehlman

et al. 2017

ACP

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BackgroundSpectrins are the principal skeletal proteins involved in cellular adhesion, migration, and signal transduction, and have been shown to be associated with integrate structure and function in complex tissue of all multi cellular organisms. The present study investigates the role of spectrins in metastasis. MethodsThree categories of samples were studied. First, spectrin expression patterns were examined in 66 normal, benign and malignant samples. Second, 188 primary tumor cases, including 69 counts of metastasis, were studied. Finally 29 cases of body fluid metastasis without available primary tumor samples were studied. Expression of spectrin isoforms αII, βI, βII, and βIII were studied by immunohistochemistry in primary tumors (188 cases) with lymph node, remote organ, and body fluid metastasis; the 26 cases of malignant body fluids were paired with their corresponding primary tumors and analyzed. ResultsIn this paper we focus on the expression of αII and βIII isoforms. We found an aberrant loss of αII mainly in breast invasive ductal carcinoma, among other cancers. Positive body fluids had markedly reduced αII and βIII expression, (P<0.0009, P<0.0001 respectively). βII remained present in all the primary tumors and cytology samples. No difference was identified in lymph node or remote organ metastasis. ConclusionWe posit based on the established role of spectrins in various forms of cancer and in cell structure and adhesion that spectrins play an important role in maintaining tissue structure, with aberrant expression associated with body fluid metastasis. Subsequently, a panel of 188 cases of primary carcinoma was selected for analysis. Cases included colon (N=77), stomach (N=3), breast (N=9), endometrium carcinoma (N=3), ovary carcinoma (N=3), lung carcinoma (N=91, adenocarcinoma N=76 and squamous cell carcinoma N=15), and thyroid (N=2). 69 counts of metastasis were included within this cohort of 188 carcinomas, as follows: regional positive lymph nodes (N=33) from colon and breast; remote organ metastasis (N=10) from colon, thyroid, and breast, to lung and liver; and malignant body fluids (N=26), primarily from the colon, stomach, liver, breast, uterine endometrium, ovary, and lung. These 26 cases of malignant body fluids were paired with their corresponding primary tumors and analyzed. Some of these 69 metastatic samples represented single patients with multiple forms of metastasis; thus, this count of 69 metastasis samples does not equal 69 individual cases, as one case can have more than one metastasis. Keywords Tumor microarraysFor larger tissues, tumor microarrays were constructed from formalin-fixed, paraffin-embedded samples using a manual tissue arrayer (Chemicon International Temecula, CA). To ensure adequate sampling, each case was represented in triplicates using 1.0mm cores. For smaller biopsy samples, histologic contiguous sections were individually prepared. The immunohistochemical staining conditions for spectrin isoforms αI, αII, βI, βII and βIII are the same as previousl...

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Spectrin-anchored phosphodiesterase 4D4 restricts cAMP from disrupting microtubules and inducing endothelial cell gap formation

Cited by 51 publications

References 62 publications

Role of tau protein on neocortical and hippocampal oscillatory patterns

Role of tau protein on neocortical and hippocampal oscillatory patterns

Protein kinase A phosphorylation of tau-serine 214 reorganizes microtubules and disrupts the endothelial cell barrier

Loss of αII and βIII Spectrin Isoforms in Body Fluids is Associated with Metastasis

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